Glycolysis

Lecture 3
2nd year rehabilitation
Objectives:
1. Describe the process of glycolysis, and breakdown of glucose in
various cells of the body.

2. list the steps of glycolysis

3. Calculate the energy derived from glycolysis

4. Explain the regulation of glycolysis

5. Discuss the diseases caused by pyruvate kinase diffecincy

 Carbohydrate Metabolism
• It is mainly Glucose metabolism which has many anabolic and
catabolic pathways:

• Major pathway for Glucose oxidation:

– Glycolysis
– Citric Acid Cycle

• Minor Pathways for Glucose oxidation:

– HMP shunt

• Glycogenesis: glycogen formation

• Glycogenolysis : glycogen degradation

• Gluconeogenesis: synthesis of glucose from non carbohydrate

sources
 Glycolysis
Glycolysis is the sequence of 10 reactions that break down one
molecule of glucose into:
–2 molecules pyruvic acid (3C) (in presence of O2)
– OR 2 molecules of lactic acid (in absence of O2).
– with the generation of 2 ATP & 2 NADH.

Site:
– In the cytoplasm of all cells of the body
Two types of glycolysis: (aerobic and anaerobic)

1. Aerobic glycolysis in the presence of O2

2. Anaerobic glycolysis in the absent of O2
Steps of glucose oxidation:
1 Glucose
Aerobic
Glycolysis
( in Cytoplasm )

O2 absent (Anaerobic)

2 Pyruvic Acid 2 Lactic Acid

O2 present (Aerobic)

TCA cycle
(In mitochondria) Aerobic Phase
O2 present

6 CO2 + H2O
Function of glycolysis:
1. In the tissue:
Glycolysis is the only energy producing pathway for many
tissues when oxygen is limited e.g.
– RBCs as they lack mitochondria
– Exercising skeletal muscles

2. In the brain:
Glycolysis is a major source of energy in the brain. Glucose is
the main fuel for the brain (it uses about 120 gm/day), partially
breaking glucose to be oxidized completely by Kreb’s cycle.

3. Formation of 2, 3 biphosphoglycerate in RBCs.

 Steps of glycolysis
Steps of glycolysis can be classified into two phases:
I- Phase one: Energy Investment
– Require 2 moles of ATP
– [Glucose gives 2 moles of glyceraldehyde-3-P].

II- Phase two: Energy yielding

In this phase the 2 moles of gIyceraldehyde-3- P are converted into:
– 2 moles of pyruvate (aerobic) OR
– 2 moles of lactate (anaerobic).
– ATP and NADH + H+ are produced
Phase I

dehydrogenase

Phase II
Steps of phase 1
1. The first step in the metabolism of glucose after entering a cell
is phosphorylation.
•phosphorylation is catalyzed by Hexokinase in extrahepatic
tissues or Glucokinase in liver.
•It requires ATP molecule.
•It is irreversible
•Importance of phosphorylation of glucose:
•Phosphorylation of sugars prevents their leakage from the
cell.
•Phosphorylation of sugars facilitates their binding to active
sites of enzymes.
Phase 1
 Differences between hexokinase and glucokinase enzymes:

Hexokinase Glucokinase
Site Extra-hepatic tissues Liver

Km Low 2mg% High 200mg%

Velocity of reaction low high

Inhibition By G 6-P No inhibition

Induction by insulin Not inducible inducible
 Importance of the difference between
glucokinase and hexokinase

The difference in Km and Vmax allows:

– The liver glucokinase to act when intracellular glucose level is high
(after carbohydrate meals) to remove glucose rapidly from portal
blood and prevent large amount of it from reaching systemic
circulation.

– Tissue hexokinase to use glucose when blood glucose level decrease.

Allosteric inhibition of hexokinase by its product (G6P) allows:

• Peripheral tissue to use glucose only when it is required to supply
energy.
• Liver glucokinase which is not inhibited by its product continue to
accumulate glucose and store it as glycogen
2- Phosphohexose Isomerase reaction:
• Glucose 6- Phosphate isomerase Fructose 6- Phosphate

3- Phosphofructokinase-1, (PFK-1)
• Fructose 6- Phosphate Phosphofructokinase-1 fructose 1,6-bisphosphate

ATP ADP
• This reaction is irreversible.
• This is the most important step in regulation of glycolysis.
4- Aldolase
• fructose 1,6-bisphosphate Aldolase dihydroxyacetone phosphate + glyceraldehyde 3-P

5- Triose Phosphate Isomerase:

• dihydroxyacetone phosphate Isomerase glyceraldehyde 3-P
Phase 2
 Steps of phase 2
6- Glyceraldehyde-3-Phosphate Dehydrogenase
Glyceraldehyde-3-Phosphate Dehydrogenase 1,3- bisphosphoglycerate
Pi NAD+ NADH + H+

• This reaction is reversible.

• Fate of NADH produced:

– In aerobic glycolysis, NADH is oxidized in respiratory chain
resulting in synthesis of 3 ATP molecules
– In anaerobic glycolysis, NADH is oxidized in reaction catalyzed by
lactate dehydrogenase.
7- Phosphoglycerate Kinase

1,3-bisphosphoglycerate Kinase 3-phosphoglycerate

ADP ATP
• The high-energy phosphate of 1,3-BPG is used to form ATP
(substrate level phosphorylation)
• This reaction is reversible.
In erythrocytes:

ATP

Importent for hemoglobin affinity to o2

N.B.
In erythrocytes, there is an associated pathway in which 1,3 biphosphoglycerate is
converted to 2,3BPG
2,3 BPG is an important regulator of hemoglobin's affinity for oxygen. Note that
2,3-bisphosphoglycerate phosphatase degrades 2,3BPG to 3-phosphoglycerate, a
normal intermediate of glycolysis without production of ATP.
8. Phosphoglycerate Mutase:
3-phosphoglycerate mutase 2- phosphoglycerate
– This reaction is reversible
9. Enolase
2- phosphoglycerate Enolase phosphoenolpyruvate

10- Pyruvate Kinase

• phosphoenoylpyruvate pyruvate kinase pyruvic acid

ADP ATP
• In this reaction, ATP is formed (substrate level phosphorylation)
• This reaction is irreversible
Reduction of Pyruvate to lactate (Anaerobic glycolysis)

In RBCs (due to lack of mitochondria) and in exercising muscle

(due to lack of oxygen), NADH produced is oxidized in a reaction
catalyzed by lactate dehydrogenase which reduces pyruvate into
lactate.

RBC
The Energy derived from glycolysis
 The Energy derived from glycolysis
Phase 1: Phase 2:
Energy Energy
investment Generation

- Needs 2 ATP - Makes 4 ATP

- HexoKinase - 2 by Phosphoglycerate kinase
- Phosphofructokinase - 2 by Pyruvate kinase

4-2=2
- Aerobic glycolysis = 2 ATP + 6 ATP ( from 2 NADH
oxidation in ETC in the mitochondria ) = 8 ATP
-Anaerobic glycolysis = 2 ATP
 Aerobic glycolysis Anaerobic glycolysis

Site - Under aerobic conditions - Under anaerobic conditions e.g.

in most tissues decrease oxygen supplies
(exercising muscles) or absence
of mitochondria (RBCs)

End Product - Pyruvic a. - Lactic a.

Fate of NADH Oxidized in respiratory - Oxidized by enzyme lactate

chain dehydrogenase

Energy yield 2 ATP + 2 NADH (6ATP) - 2 ATP

= 8 ATP
 Pyruvate kinase (PK) deficiency
Genetic deficiency
• Leads to haemolytic anemia WHY?

• Because RBCs depend completely on

glycolysis as a source of energy

• With decrease activity of PK so no enough

ATP will produce

• In addition membrane integrity is effected

• Death and lysis of RBCS

• Hemolytic anemia
 Regulation of glycolysis
• Regulation of glycolysis occur in the three irreversible
reactions (reactions 1, 3, 10)
• Hexokinase is allosterically inhibited by its product (glucose 6
phosphate)
• Phosphofructokinase 1: this is the most important regulatory
enzyme, it is regulated in two ways:
– Energy level of the cell; high ATP level allosterically inibit the
enzyme, high level of AMP allosterically activates the enzyme.
– Fructose 2,6 biphosphate (increase by insulin) allosterically
activate the enzyme
• Pyrvate kinase:
– Covalent modification: phosphorylation of the enzyme (by
hormone glucagons) inhibit the enzyme

• Hormonal regulation: (Induction & Repression of enzymes)

– Insulin induces (increases synthesis) of glucokinase,
phosphofructokinase and pyruvate kinase
– Glucagone represses (decreases snthesis) of these enzymes.
 Regulation of Glycolysis
3 irreversible reaction

Reaction 1 Reaction 3 Reaction 10

Hexokinase Phosphofructokinase Pyruvate kinase

-Inhibited by G6P -Phosphorylation of the
enzyme will inhibits it
(Allosteric inhibition) Most imp -P by glucagon
-(Covalent modification)

Energy Levels Fructose 2,6 bisphosphate

Increase by insulin
(Allosteric inhibition) High ATP x (inhibit) (Allosteric activation)
(Allosteric activation) High AMP✔️ (Activate)
Phase I

1st irreversible reaction

2nd irreversible reaction

dehydrogenase

Phase II

3rd irreversible reaction

 Oxidation Of Extra-Mitochondrial NADH

-NADH is produced in glycolysis in the cytosol

- NADH Cannot pass through the mitochondria membrane

to reach the ETC in the mitochondria

- So it Needs shuttle

-Two shuttles can function in the transfer of hydrogen from

NADH in the cytosol to mitochondria:
Glycerophosphate Shuttle
Malate - Aspartate Shuttle
 Malate-Aspartate Shuttle
Glycerophosphate Shuttle
 Recommended reading

• Lippincott’s illustrated reviews

biochemistry